Determination of binding affinities using hyperpolarized NMR with simultaneous 4-channel detection

Journal of Magnetic Resonance
Yaewon KimChristian Hilty

Abstract

Dissolution dynamic nuclear polarization (D-DNP) is a powerful technique to improve NMR sensitivity by a factor of thousands. Combining D-DNP with NMR-based screening enables to mitigate solubility or availability problems of ligands and target proteins in drug discovery as it can lower the concentration requirements into the sub-micromolar range. One of the challenges that D-DNP assisted NMR screening methods face for broad application, however, is a reduced throughput due to additional procedures and time required to create hyperpolarization. These requirements result in a delay of several tens of minutes in-between each NMR measurement. To solve this problem, we have developed a simultaneous 4-channel detection method for hyperpolarized 19F NMR, which can increase throughput fourfold by utilizing a purpose-built multiplexed NMR spectrometer and probe. With this system, the concentration-dependent binding interactions were observed for benzamidine and benzylamine with the serine protease trypsin. A T2 relaxation measurement of a hyperpolarized reporter ligand (TFBC; CF3C6H4CNHNH2), which competes for the same binding site on trypsin with the other ligands, was used. The hyperpolarized TFBC was mixed with trypsin and the ligan...Continue Reading

Citations

Apr 5, 2019·Angewandte Chemie·Jasmine Viger-GravelLyndon Emsley
Feb 5, 2019·Journal of Medicinal Chemistry·Yunyi Wang, Christian Hilty
Sep 5, 2021·Progress in Nuclear Magnetic Resonance Spectroscopy·Ēriks KupčeTim D W Claridge

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